LOCUS       sequence-373229-        7955 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of SRGAP1-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7955)
  AUTHORS   Liang X, Budnar S, Gupta S, Verma S, Han SP, Hill MM, Daly RJ, 
            Parton RG, Hamilton NA, Gomez GA, Yap AS
  TITLE     Tyrosine dephosphorylated cortactin downregulates contractility at 
            the epithelial zonula adherens through SRGAP1.
  JOURNAL   Nat Commun. 2017 Oct 5;8(1):790. doi: 10.1038/s41467-017-00797-w.
  PUBMED    28983097
REFERENCE   2  (bases 1 to 7955)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2017-10-5"; volume: "8(1):790. doi"; pages: " 
            10.1038/s41467-017-00797-w"
COMMENT     Sequence Label: pEGFP-SRGAP1
FEATURES             Location/Qualifiers
     source          1..7955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        70..373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        374..577
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     528..548
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3904..3913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3910..3912,3913..3915,3916..4629)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3910..3912,3913..3915,3916..4626)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3955..3976)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4216..4235)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4563..4584
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4752..4873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4789..4808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4843..4862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4880..5335)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5017..5038)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5229..5248
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5362..5466
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5468..5825
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5468..5659
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5491..5511)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5676..5811
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5860..6654
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5860..6654
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5914..5933)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6524..6543
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6842..6861)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6886..6933
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7262..7850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7751..7770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
       61 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      121 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      181 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      241 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      301 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      361 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      421 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      481 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      541 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
      601 ctagcgctac cggactcaga tctcgaggcc accatgtcca ccccgagccg attcaagaag
      661 gacaaagaga tcatagccga gtatgaaagt caagtcaaag aaattcgagc tcaactggta
      721 gaacaacaaa aatgcctgga gcagcaaacg gagatgcgag ttcagcttct ccaggatctg
      781 caagatttct tccgaaaaaa agctgaaatt gagacggaat attcccggaa tctagagaag
      841 ttagcagaaa ggttcatggc aaaaacaaga agcactaagg atcatcaaca atacaagaaa
      901 gaccagaacc tgttgtctcc agtgaactgc tggtatttgc tcctgaacca agtaaggaga
      961 gaaagcaaag accatgcaac cttgagtgac atctatctga acaatgtgat tatgcggttc
     1021 atgcagataa gtgaggattc taccaggatg tttaaaaaga gcaaagagat tgcattccaa
     1081 cttcatgagg atttaatgaa ggttcttaat gagctttata cggtgatgaa aacataccat
     1141 atgtatcatg cagagagcat cagtgcagag agcaagctga aagaggccga aaaacaagag
     1201 gaaaagcaaa ttgggagatc tggtgatcca gtcttccata ttcgactaga ggagagacat
     1261 caacggcgaa gctctgtaaa gaaaattgaa aaaatgaaag aaaaaagaca agcaaaatat
     1321 tcagaaaata agctaaaatc aattaaggca cggaacgaat atctcctaac acttgaagcc
     1381 accaatgcct cagttttcaa gtactatatt catgatcttt ctgatttaat tgattgctgt
     1441 gatcttggct accatgcaag tctgaacaga gccctaagaa catatctgtc tgcggagtac
     1501 aaccttgaaa cctccagaca tgagggctta gacattattg agaatgcagt tgataattta
     1561 gagcccagga gcgataagca gagattcatg gagatgtacc ctgctgcgtt ctgtccacca
     1621 atgaagtttg agtttcagtc tcacatgggt gatgaggtgt gccaggtcag tgcccagcag
     1681 ccagtccagg cagagctcat gctcaggtac caacagttgc agtcccgcct tgccacgctc
     1741 aaaatcgaga atgaagaggt taagaaaacg actgaagcca ccttgcagac gatacaagat
     1801 atggtcacca tcgaggacta tgatgtttct gaatgcttcc agcacagtcg ttccacagaa
     1861 tctgtgaagt ccactgtctc tgaaacctac ctgagtaaac ccagcatcgc caagagaaga
     1921 gccaaccagc aggaaactga acagttctac ttcatgaaac tcagagaata tttggaaggc
     1981 agtaatctca tcacaaaact tcaagccaaa catgacttgc tgcagaggac cctgggagaa
     2041 ggtcatagag ctgaatatat gactacaagg cctccaaatg ttccccctaa gccccagaaa
     2101 cacaggaagt ccaggccccg ctcacagtat aatactaagt tgtttaatgg ggatttggaa
     2161 acattcgtca aggactcagg acaggttatt cccctcattg tggaaagctg tattcggttc
     2221 atcaatctct atggtcttca gcatcagggg attttcagag tgtctggttc ccaggtggaa
     2281 gtcaatgata ttaaaaattc atttgagaga ggtgaaaatc ctttggctga tgaccagagt
     2341 aaccatgata ttaactcagt tgctggcgtt ctgaagctct atttccgtgg gctggaaaac
     2401 cccctctttc ctaaggaaag atttaacgat ctgatttctt gtatcagaat agataatctc
     2461 tatgagaggg cgcttcacat ccgcaaactc ctcctgactt tgcccaggtc ggtccttata
     2521 gtgatgaggt acctctttgc cttcctcaat catctatcac agtacagcga tgagaatatg
     2581 atggaccctt ataacctggc catttgcttt ggcccaacat tgatgcctgt cccagaaata
     2641 caggatcaag tgtcttgcca ggcacatgtg aatgaaatta tcaaaaccat catcatccac
     2701 catgagacta ttttcccaga tgctaaagag ctggatggcc ctgtttatga gaaatgtatg
     2761 gctggagatg actattgcga cagcccatac agtgagcacg gtacattgga ggaagtggac
     2821 caagatgctg gtacagagcc ccacacaagt gaagatgaat gtgagccaat agaagcaata
     2881 gccaagtttg actatgttgg gcggtccgcc agagaactat ccttcaagaa gggtgcctcc
     2941 ctgctgctgt atcaccgtgc atctgaggac tggtgggaag gcaggcacaa cgggattgac
     3001 gggctggtgc ctcaccagta tatagtggtg caggatatgg atgatacgtt ttcagacact
     3061 ctgagccaaa aagccgacag tgaggccagc agtgggccag tcacggaaga caagtcctca
     3121 tccaaggaca tgaactcccc gacagaccgt catcctgacg gctatttagc caggcaacga
     3181 aaaagaggag agccaccccc tccagtaagg cgtcctggca ggaccagtga tggccattgc
     3241 ccgctccacc ctccacatgc cctttctaac tcctcagttg acctagggtc cccaagcctt
     3301 gccagtcacc cccggggcct gctgcagaac cgtggcctca acaatgacag tcctgagcgg
     3361 aggcgcaggc ctggccatgg cagcctgacc aacatcagcc ggcacgactc cctcaagaag
     3421 atcgacagcc ctcccattag aaggtccacg tcatcagggc aatacacggg cttcaatgac
     3481 cacaagccac tggacccaga gacaattgct caggatattg aagaaacgat gaacacagct
     3541 ttgaatgaac tccgagaact ggagagacag agcacagcaa agcatgcccc tgatgtggtg
     3601 ctggataccc tggagcaagt gaaaaactct cccacccctg ccacttccac ggaatctctc
     3661 agccctttgc acaacgttgc cctcaggagc tccgagcctc agattcgacg tagcacgagc
     3721 tcctccagtg acacaatgag tactttcaag cctatggtgg cacccagaat gggcgtgcag
     3781 ctgaagcctc cagcccttag gccaaaacct gctgttcttc caaaaacaaa tcctaccata
     3841 ggacctgccc cacctcccca gggtccaaca gacaagtcat gcacaatggg ggatccaccg
     3901 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3961 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4021 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     4081 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     4141 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4201 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     4261 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4321 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     4381 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     4441 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4501 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     4561 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     4621 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     4681 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     4741 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4801 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4861 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4921 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4981 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5041 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     5101 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     5161 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     5221 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5281 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     5341 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5401 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5461 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5521 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5581 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5641 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5701 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5761 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5821 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5881 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5941 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6001 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     6061 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6121 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6181 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6241 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6301 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6361 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     6421 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6481 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6541 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6601 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6661 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6721 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     6781 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     6841 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     6901 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     6961 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     7021 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     7081 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     7141 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7201 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7261 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7321 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7381 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7441 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7501 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7561 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7621 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7681 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7741 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7801 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7861 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7921 cgttatcccc tgattctgtg gataaccgta ttacc
//
